Morphological, physiological and bi

Morphological, physiological and biochemical character- istics were determined by using previously described methods (Endo & Okada, 2005), except that API 50 CHL galleries (bioMe ́rieux) were used for the determination of acid production from carbohydrates. FYP broth was used as a basal medium. The detailed characteristics for the novel isolates are given in the species description and are compared with the characteristics of their phylogenetic neighbours L. fructivorans, L. homohiochii, L. lindneri and L. sanfranciscensis in Table 1. From the 49 carbohydrates tested, the new isolates produced acids from only D-glucose and D-fructose. To the authors’ knowledge, this result represents the lowest number of fermented carbohydrates thus far described for LAB. D-Fructose was fermented within 24 h, but D-glucose was fermented after 48 h. The novel isolates could be distinguished from their phylogen- etic relatives by their acid production patterns from carbohydrates (Table 1). The new isolates grew well at acidic to weak alkaline pH (4.0–7.5), whereas their phylogenetic relatives were acidophilic. Growth at pH 6.5–7.5 is thus a key characteristic that differentiated the new isolates from phylogenetically closely related species. Catalase activity was not detected from the three strains grown in FYP broth under aerobic conditions but was detected when the novel strains were cultured on FYP agar containing 5 % (v/v) sheep blood under aerobic conditions. The strains produced DL-lactic acid, ethanol and acetic acid at a ratio of 1 : 0.8 : 0.2 (Endo et al., 2009). The three strains reached optical density values (660 nm) of 1.0 after 2 days incubation in the presence of 200 g l21 D-fructose. The novel strains took 3–4 days to reach optical density values of 1.0 in the presence of 300 g l21 D-fructose.